ACE2 and AT4R are present in diseased human blood vessels.

A growing body of evidence suggests that the angiotensin II fragments, Ang(1-7) and Ang(3-8), have a vasoactive role, however ACE2, the enzyme that produces Ang(1-7), or AT4R, the receptor that binds Ang (3-8), have yet been simultaneously localised in both normal and diseased human conduit blood vessels. We sought to determine the immunohistochemical distribution of ACE2 and the AT4R in human internal mammary and radial arteries from patients undergoing coronary artery bypass surgery. We found that ACE2 positive cells were abundant in both normal and diseased vessels, being present in neo-intima and in media. ACE2 positive immunoreactivity was not present in the endothelial layer of the conduit vessels, but was clearly evident in small newly formed angiogenic vessels as well as the vaso vasorum. Endothelial AT4R immunoreactivity were rarely observed in either normal and diseased arteries, but AT4R positive cells were observed adjacent to the internal elastic lamine in the internal mammary artery, in the neo-intima of radial arteries, as well as in the media of both internal mammary artery and radial artery. AT4R was abundant in vaso vasorum and within small angiogenic vessels. Both AT4R and ACE2 co-localised with smooth muscle cell alpha actin. This study identifies smooth muscle cell alpha actin positive ACE2 and AT4R in human blood vessels as well as in angiogenic vessels, indicating a possible role for these enzymes in pathological disease.

Calcitonin receptor immunoreactivity associated with specific cell types in diseased radial and internal mammary arteries.

AIMS: To determine and quantify calcitonin receptor (CTR) immunoreactivity associated with specific cell types within, and associated with, the endothelial layers, neo-intima, media and vasa vasorum of diseased radial and internal mammary arteries. METHODS AND RESULTS: Immunohistochemistry and anti-CTR antibodies were used to identify positive cells within remnants of diseased human radial (n = 3) and internal mammary arteries (n = 4) that remained after bypass surgery. Three cell types expressed CTR, including endothelial cells, fibroblast-like cells within the neo-intima, and cellular structures aligned with the smooth muscle cells of the media. Other smaller cells within the surrounding parenchyma of the vasa vasorum of diseased vessels and blood-borne cells were also immunoreactive. Immunoquantification of CTR expression (Intensity x Proportional Area) in the endothelium (P < 0.05), neo-intima (P < 0.02) and media (P < 0.03) established a significant statistical correlation (Students' two-tailed t-test) with the ratio of intimal/media thickness. CONCLUSIONS: Increased immunoreactivity developed using anti-CTR antibodies was associated with specific cell types in the endothelial layers, neo-intima, media and vasa vasorum of diseased regions of radial and internal mammary arteries, in which there was an increased intimal/media ratio. Furthermore, CTR+, blood-borne cells present in the vessels of diseased regions suggest recruitment into these surrounding tissues.

Preoperative assessment of hand circulation by means of Doppler ultrasonography and the modified Allen test.

OBJECTIVE: The aims of this study were as follows: (1) to evaluate Doppler ultrasonography in assessing hand collateral circulation; (2) to define the criteria for an abnormal Doppler ultrasonography dynamic test result; and (3) to validate the modified Allen test. METHODS: The hand circulation of 71 patients scheduled for coronary artery bypass grafting was assessed by means of the Allen test and Doppler ultrasonography. The flow in the superficial palmar branch of the radial artery, the ulnar artery, and the dorsal digital thumb artery with and without radial artery compression were recorded. Flow patterns in the superficial palmar branch of the radial artery, the ulnar artery, and the dorsal digital thumb artery with radial artery compression were categorized into 4 groups: (1) no flow; (2) decreased flow; (3) reversed flow; and (4) increased flow. RESULTS: Among the 71 hands, 4 (5.6%) had an abnormal Allen test result (>10 seconds). Seven (10.6%) of 66 superficial palmar branches of the radial artery, 3 (4.2%) of 71 ulnar arteries, and 2 (2.8%) of 71 dorsal digital thumb arteries showed no flow with radial artery compression, as measured by Doppler ultrasonography. There were significant differences among the 4 groups (superficial palmar branch of the radial artery: F = 7.0, P <.001; ulnar artery: F = 13.1, P <.001; and dorsal digital thumb artery: F = 8.4, P <.001) for the Allen test. Pairwise comparisons showed that when subjected to an Allen test, category 1 patients (no flow) had significantly longer recovery times compared with the other groups (P <.02 in all cases) for the superficial palmar branch of the radial artery, the ulnar artery, and the dorsal digital thumb artery. CONCLUSION: Absence of flow in the dorsal digital thumb artery with radial artery compression is considered an absolute contraindication to radial artery harvesting. An increased recovery time with the modified Allen test predicts absence of flow in the dorsal digital thumb artery in Doppler ultrasonographic flow patterns. This demonstrates the validity of the modified Allen test for primary screening.

Cardiac rehabilitation and secondary prevention.

Group programs of cardiac rehabilitation and secondary prevention provide better outcomes than medical consultations alone. Tightly specified goals for secondary prevention are now more rigorous than ever before and should be reached by all patients. After acute myocardial infarction, patients require beta-blockers, aspirin, lipid-lowering agents and angiotensin-converting enzyme inhibitors. Psychosocial adjustment problems are common in cardiac patients and their families, but these can be significantly reduced by appropriate rehabilitation strategies. Patients with additional needs should be identified (e.g., some working patients require work assessment, employer contact, additional exercise and specified return-to-work guidelines).

Construction of a novel oncogene based on synthetic sequences encoding epidermal growth factor.

The autocrine model postulates that constitutive release of a mitogenic growth factor can lead to uncontrolled proliferation and cell transformation. A synthetic polynucleotide encoding epidermal growth factor conferred a tumorigenic phenotype on cells. These cells were transformed through the action of an autocrine circuit having an extracellular component.

Regulated high-level expression of the Herpes simplex type I thymidine kinase gene in Escherichia coli.

A plasmid-borne Herpes simplex virus type 1 (HSV-1) thymidine kinase (TK) gene (tk) was expressed in Escherichia coli by inserting a 203-bp lacL8/UV5 promoter-operator segment, in frame, 53 bp 5' to the native tk translational start codon. The hybrid gene created by this fusion encodes a polypeptide which has 25 additional amino acids on the amino terminus of the HSV-1 TK protein and phenotypically complements a tdk- mutation of E. coli. This fusion polypeptide has been characterized by maxicell, immunoprecipitation, and native gel techniques, and its activity is inhibited by anti-HSV-1 antibody. In a tk expressor strain containing a F' lacIq (which overproduces the lactose repressor), the isopropyl-beta-D-thiogalactoside (IPTG) causes greater than 1000-fold coordinate induction of the plasmid-encoded TK and chromosomal beta-galactosidase activities. Pulse-chase induction demonstrates the fused TK polypeptide to be as stable as beta-galactosidase. HSV-1 tk-specific RNA isolated from this bacterial strain has a short half-life characteristic of bacterial messages.